Stimulatory effect of ipriflavone on formation of bone-like tissue in rat bone marrow stromal cell culture

The effects of ipriflavone (IP) (10^–5 M) on bone formation were studied in stromal cells from the femoral bone marrow of young adult rats cultured for 21 days in the presence of -glycerophosphate and dexamethasone. Stereoscopic microscopy showed nodule formation after 14 days of culturing, and both the number and the size of the nodules increased with time. The alizarin-red-stained calcified area in the nodules in the IP group was nearly 4 times as large as that in the control after 21 days. Light and electron microscopy revealed the presence of many osteoblast-like cells with developed rough endoplasmic reticulum and Golgi apparatus in the nodules in the control group after 14 days, and a collagenous fibril network was seen among the cells. After 21 days, calcification of the dense collagenous fibril network and bone matrix-like tissue were observed in many nodules, resulting in the formation of bone-like tissue containing osteocyte-like cells. In the IP group, the collagenous fibril network area in the nodules was greater than that in the control after 14 days, and a further increase in both the dense collagenous fibril network area and calcified bone-like tissue area was observed after 21 days. These findings indicate that IP stimulates bone-like tissue formation in the rat bone marrow stromal cell culture, suggesting that the promotion of collagen production by osteoblasts is involved in the stimulation of bone-like tissue formation by IP.     

The putative mechanism of thrombosis in antiphospholipid syndrome: impairment of the protein C and the fibrinolytic systems by monoclonal anticardiolipin antibodies

The mechanism of thrombosis in patients with antiphospholipid syndrome is not clear. To investigate it, we examined the effect of monoclonal anticardiolipin (aCL) antibodies and beta2-glycoprotein I (beta2-GPI), which is required for formation of the aCL epitopes, on activated protein C (APC) and on fibrinolytic activity. First, APC activities were measured in the presence and absence of beta2-GPI or gamma M immunoglobulin (IgM) monoclonal aCLs (EY1C8 and EY2C9), or both, established from peripheral blood lymphocytes obtained from a patient with aCL. beta2-GPI exhibited a procoagulant activity by inhibiting APC activity as well as an anticoagulant activity by inhibiting thrombin generation. Any further inhibition of APC activity was caused by monoclonal aCL, and then only in the presence of beta2-GPI. The remaining tissue plasminogen activator (t-PA) of the sample consisting of beta2-GPI, two-chain recombinant t-PA, and plasminogen activator inhibitor (PAI)-1 was measured by a chromogenic assay using the synthetic substrate S-2251, Glu-plasminogen, and soluble fibrin monomer. beta2-GPI protected t-PA activity from inhibition by PAI-1. However, monoclonal aCLs (EY1C8 and EY2C9) inhibited the effect of beta2-GPI on fibrinolytic activity; that is, monoclonal aCLs inhibited fibrinolytic activity by elevating PAI-1 activity. Thrombosis in patients with aCL can be explained in part by both the inhibition of APC anticoagulant activity and the impairment of fibrinolytic activity by aCL.     

Autoantibody against erythrocyte protein 4.1 in a patient with autoimmune hemolytic anemia

We observed the presence of a new autoantibody, anti-erythrocyte protein 4.1, in a patient with autoimmune hemolytic anemia (AIHA). Western blotting analysis revealed that IgG from the patient's plasma reacted with erythrocyte protein 4.1. However, among other patients with hemolytic diseases (six having AIHA and three each having either hereditary spherocytosis, elliptocytosis, or lead poisoning) as well as among control subjects, no antibody activity to protein 4.1 was observed. In addition to the anti-protein 4.1 antibody, two different kinds of anti-erythrocyte antibodies were detected by conventional serological studies in this patient. One of them was an anti-Ena-like antibody in the eluate from the patient's erythrocytes, while another was the anti-S-specific antibody in the plasma. An elution study and an absorption study using S antigen-positive erythrocytes demonstrated that the anti-protein 4.1 antibody differed from both the anti-Ena-like antibody and the anti-S antibody. Familial analysis of the patient revealed the same antibody in her brother, who did not have hemolytic anemia. These results demonstrate that anti-protein 4.1 antibody is considered to be included in the spectrum of anti-cytoskeleton autoantibodies, which have been observed in patients having increased cell lysis as well as in healthy subjects.     

Rapid and simple determination of epoxyeicosatrienoic acids in rabbit renal artery by reversed-phase HPLC with fluorescence detection

A liquid chromatographic method with fluorescence detection coupled with a solid-phase extraction was applied to the rapid determination of epoxyeicosatrienoic acids (EETs) in the rabbit renal artery. The EETs were extracted with an acetonitrile from renal artery homogenate and concentrated by a solid-phase extraction method. The concentrated EETs were reacted directly with a 6, 7-dimethoxy-1-methyl-2 (1H)-quinoxalinone-3-propionyl-carboxylic acid (DMEQ) hydrazide and separated by a reversed-phase HPLC with eluting a combination of a step-wise and a gradient of a mixture of methanol and water. The content of EETs in the renal arteries was significantly greater in the 0.5% cholesterol fed rabbits than in control rabbits. It is suggested that hyperchlesterolemia increases the production of EETs in the rabbit renal artery.     

CD30-positive anaplastic large cell lymphoma of bone treated with autologous peripheral blood stem cell transplantation

A 30-year-old man was admitted to our hospital with the chief complaints of fever and pain in the right shoulder, axilla, and inguinocrural region. Computed tomography, magnetic resonance imaging, 67Ga-scintigraphy, and bone-scintigraphy revealed osteolytic lesions in the ribs and right ilium. Biopsy specimens from lesions in the right ilium confirmed the diagnosis of CD 30-positive anaplastic large cell lymphoma. The patient was treated with 6 courses of CHOP therapy followed by high-dose chemotherapy and autologous peripheral blood stem cell transplantation. He achieved and remained in remission with no evidence of relapse 14 months later.     

The effectiveness of diverse technology-based instructions in assisting people with Alzheimer’s disease with medication management

Abstract

Purpose: In patients with mild cognitive impairment and Alzheimer’s disease (AD), faculties associated with instrumental activities of daily living (IADL) decline owing to reduced cognitive function. One type of IADL is medication behaviour. Medication management is critical for patients with AD. Previous studies have reported that presentations using speech and images are effective for facilitating IADL autonomy but there are few examinations of effective presentation methods. Therefore, we examined what kind of display methods are effective in helping patients with AD with medication management.

Materials and methods: Ten healthy elderly and 9 patients with mild AD were asked to perform the task of taking out medicine bags from a case at a designated time. We gave 3 kinds of instructions and examined the differences in participants’ reaction times. Task 1 included verbal instructions alone, Task 2 included verbal instructions and pictorial and written instructions, and Task 3 used a video conference system (presenting pictorial, written, and verbal instructions) at a designated time. Task 3 could be conducted remotely over the internet. The relationship between these results and neuropsychological tests was also explored.

Results: Task 3 was an effective method for patients with mild AD. In addition, we found correlations between the methods of Tasks 1–3 and the Japanese version of the Mini-Mental State Examination (MMSE-J).

Conclusions: The method of Task 3 may lead to home support for patients with AD. The MMSE-J could be used to identify changes in the adaptive functioning of patients exposed to distinct presentation methods.

• Implications for rehabilitation

• Appropriate transmission methods will increase the IADL autonomy of patients with mild AD who have memory impairment. While still images alone are not effective for patients with mild AD, when combined with verbal instructions, they prove effective for this group.

• The results of this study are useful for providing patients with mild AD with support in their IADLs, especially when methods that use images plus speech are employed. As participants were instructed via the internet, this study shows a way to help patients with mild AD even from a remote location.

• Until now, no studies have examined the adaptation criteria for instructive methods for patients with mild AD. This study shows that the MMSE could be used to determine the applicability of these instructive methods. The identification of cut-off values in future research could lead to more effective IADL support.

     

Development of a rapid blood transfusion system with the capability of blood purification.

Blood is usually irradiated by x-ray to prevent graft-versus-host-disease. However, plasma potassium levels of irradiated blood are rapidly increased during preservation in irradiated blood. The objectives of this study were to develop a rapid blood transfusion system for which irradiated blood can be used and to evaluate the capability of blood purification of the system. Packed red blood cells (RBC) were irradiated (15 Gy x-ray) at 21 days and preserved until 42 days after collection. A blood mixture of RBC and plasma was perfused through a dialyzer at 25, 50, 100, and 200 ml/min. Dialysate was perfused at 100, 100, 500, and 500 ml/min, respectively. Preperfusion levels of sodium, 121; potassium, 35; and chlorine, 76 mEq/L were changed to sodium, 144 to 146; potassium 2.5 to 3.0; and chlorine, 105 to 110 mEq/L, which were comparable with the levels in dialysate after perfusion for 25, 50, and 100 ml/min perfusion groups. For the 200 ml/min perfusion group, potassium was 5.3 mEq/L after perfusion which was slightly higher than other groups, but 84% of the potassium was removed by the system. Citrate levels were significantly decreased to 3.4, 28, 31, and 81 mg/dl for the 25, 50, 100, and 200 ml/min groups, respectively, after perfusions. The rapid transfusion system composed of the dialyzer and the blood pumps was effective in the removal of potassium and in the normalization of electrolytes. Irradiated blood with high levels of potassium can be safely and effectively used for this system in cases requiring massive rapid blood transfusion.